LOCUS       40924_29686        5593 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMAK705, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5593)
  AUTHORS   Hamilton CM, Aldea M, Washburn BK, Babitzke P, Kushner SR.
  TITLE     New method for generating deletions and gene replacements in 
            Escherichia coli
  JOURNAL   J. Bacteriol. 171 (9), 4617-4622 (1989)
  PUBMED    2548993
REFERENCE   2  (bases 1 to 5593)
  AUTHORS   Maples VF, Kushner SR.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2002) Genetics, University of Georgia, Life 
            Sciences Bldg., Athens, GA 30602, USA
REFERENCE   3  (bases 1 to 5593)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5593)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "1989"; volume: "171"; issue: "9"; pages: 
            "4617-4622"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUN-2002) Genetics, University of Georgia, Life Sciences Bldg., 
            Athens, GA 30602, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      487..709
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             757..1704
                     /label=Rep101(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
     promoter        complement(3191..3295)
                     /label=AmpR promoter
     promoter        3498..3600
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             3601..4257
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     primer_bind     5205..5221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5222..5278
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5291..5307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5315..5331)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5339..5369)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5384..5405)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5506..5593
                     /note="derived from vector pBR325 deposited in GenBank
                     Accession Number L08855"
ORIGIN
        1 gatccccgac agtaagacgg gtaagcctgt tgatgatacc gctgccttac tgggtgcatt
       61 agccagtctg aatgacctgt cacgggataa tccgaagtgg tcagactgga aaatcagagg
      121 gcaggaactg ctgaacagca aaaagtcaga tagcaccaca tagcagaccc gccataaaac
      181 gccctgagaa gcccgtgacg ggcttttctt gtattatggg tagtttcctt gcatgaatcc
      241 ataaaaggcg cctgtagtgc catttacccc cattcactgc cagagccgtg agcgcagcga
      301 actgaatgtc acgaaaaaga cagcgactca ggtgcctgat ggtcggagac aaaaggaata
      361 ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg
      421 ttttgtagag gagcaaacag cgtttgcgac atccttttgt aatactgcgg aactgactaa
      481 agtagtgagt tatacacagg gctgggatct attcttttta tcttttttta ttctttcttt
      541 attctataaa ttataaccac ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa
      601 tttacagagg gtctagcaga atttacaagt tttccagcaa aggtctagca gaatttacag
      661 atacccacaa ctcaaaggaa aaggactagt aattatcatt gactagccca tctcaattgg
      721 tatagtgatt aaaatcacct agaccaattg agatgtatgt ctgaattagt tgttttcaaa
      781 gcaaatgaac tagcgattag tcgctatgac ttaacggagc atgaaaccaa gctaatttta
      841 tgctgtgtgg cactactcaa ccccacgatt gaaaacccta caaggaaaga acggacggta
      901 tcgttcactt ataaccaata cgttcagatg atgaacatca gtagggaaaa tgcttatggt
      961 gtattagcta aagcaaccag agagctgatg acgagaactg tggaaatcag gaatcctttg
     1021 gttaaaggct ttgagatttt ccagtggaca aactatgcca agttctcaag cgaaaaatta
     1081 gaattagttt ttagtgaaga gatattgcct tatcttttcc agttaaaaaa attcataaaa
     1141 tataatctgg aacatgttaa gtcttttgaa aacaaatact ctatgaggat ttatgagtgg
     1201 ttattaaaag aactaacaca aaagaaaact cacaaggcaa atatagagat tagccttgat
     1261 gaatttaagt tcatgttaat gcttgaaaat aactaccatg agtttaaaag gcttaaccaa
     1321 tgggttttga aaccaataag taaagattta aacacttaca gcaatatgaa attggtggtt
     1381 gataagcgag gccgcccgac tgatacgttg attttccaag ttgaactaga tagacaaatg
     1441 gatctcgtaa ccgaacttga gaacaaccag ataaaaatga atggtgacaa aataccaaca
     1501 accattacat cagattccta cctacataac ggactaagaa aaacactaca cgatgcttta
     1561 actgcaaaaa ttcagctcac cagttttgag gcaaaatttt tgagtgacat gcaaagtaag
     1621 tatgatctca atggttcgtt ctcatggctc acgcaaaaac aacgaaccac actagagaac
     1681 atactggcta aatacggaag gatctgaggt tcttatggct cttgtatcta tcagtgaagc
     1741 atcaagacta acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa agggaaaact
     1801 gtccatatgc acagatgaaa acggtgtaaa aaagatagat acatcagagc ttttacgagt
     1861 ttttggtgca tttaaagctg ttcaccatga acagatcgac aatgtaacag atgaacagca
     1921 tgtaacacct aatagaacag gtgaaaccag taaaacaaag caactagaac atgaaattga
     1981 acacctgaga caacttgtta cagctcaaca gtcacacata gacagcctga aacaggcgat
     2041 gctgcttatc gaatcaaagc tgccgacaac acgggagcca gtgacgcctc ccgtggggaa
     2101 aaaatcatgg caattctgga agaaatagcg ctttcagccg gcaaaccggc tgaagccgga
     2161 tctgcgattc tgataacaaa ctagcaacac cagaacagcc cgtttgcggg cagcaaaacc
     2221 cgtacttttg gacgttccgg cggttttttg tggcgagtgg tgttcgggcg gtgcgcgcaa
     2281 gatccattat gttaaacggg cgagtttaca tctcaaaacc gcccgcttaa caccatcaga
     2341 aatcctcagc gcgattttaa gcaccaaccc ccccccgtaa cacccaaatc catactgaaa
     2401 gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcctcc
     2461 cgacaacaca gaccattccg tggcaaagca aaagttcaga atcaccaact ggtccaccta
     2521 caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg aggcgattca
     2581 ggcctggtat gagtcggcaa caccttcatc acgaggaagg ccccagcgct attctgatct
     2641 cgccatcacc accgttctgg tgattaaacg cgtattccgg ctgaccctgc gggctgcgca
     2701 gggttttatt gattccattt ttgccctgat gaacgttccg ttgcgctgcc cggattacac
     2761 cagtgtcagt aagcgggcaa agtcggttaa tgtcagtttc aaaacgtcca cccggggtga
     2821 aatcgcacac ctggtgattg attccaccgg gctgaaggtc tttggtgaag gcgaatggaa
     2881 agtcagaaag cacggcaaag agcgccgtcg tatctggcga aagttgcatc ttgctgtggg
     2941 gatcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3301 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     3361 aaaataggcg tatcacgagg ccctttcgtc ttcgaataaa tacctgtgac ggaagatcac
     3421 ttcgcagaat aaataaatcc tggtgtccct gttgataccg ggaagccctg ggccaacttt
     3481 tggcgaaaat gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata
     3541 agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa
     3601 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     3661 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     3721 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     3781 cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt
     3841 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     3901 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     3961 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     4021 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     4081 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     4141 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
     4201 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     4261 tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg aataagtgat
     4321 aataagcgga tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc ggttcagggc
     4381 agggtcgtta aatagccgct tatgtctatt gctggtttac cggtttattg actaccggaa
     4441 gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg
     4501 aggtaataat tgacgatatg atcatttatt ctgcctccca gagcctgata aaaacggtga
     4561 atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc
     4621 gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc
     4681 gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga
     4741 agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac
     4801 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
     4861 cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc
     4921 gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     4981 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccatg
     5041 tttgacagct tatcatcgat aagctcattc gccattcagg ctgcgcaact gttgggaagg
     5101 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     5161 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     5221 tgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg
     5281 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5341 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5401 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5461 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattggcacc attccttgcg
     5521 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     5581 aagggagagc gtc
//
